About this repo:

This repository includes all the files used to reconstruct the figures on Tabima et al. (In Prep) as well as the files not included in the submitted manuscript.

To reconstruct the figures download or clone this repository and execute the Readme.Rmd on Rstudio with the knitr and all the required packages installed. A rendered version of this document can be found on the Readme.html included in this repo

Most files are in Rmd format. This is both to create small, binary files to be executed rapidly in R and to replicate the figures as accurately as possible.

List of files:

Main Figures

  • Fig1/ (Abundance of secondary metabolite (SM) core genes in zygomycetes)
    • Zygo_SM_count.csv: CSV file summary counts of SM core genes in 69 species of zygomycetes.
  • Fig2/ (RNA expression of secondary metabolite (SM) core genes in Basidiobolus meristosporus CBS 931.37)
    • count_RNA.Rds: Rds file with TPS per gene model of Basidiobolus meristosporus CBS 931.37
  • Fig4/ (Phylogenetic origin of NRPS A-domains in Basidiobolus species)
    • Colors_arrow_origin.Rds: Rds file with color schemes for phylogenetic origin of A-domains of Basidiobolus NRPS proteins
    • Colors_arrow.Rds: Rds file with color schemes for backbone of figure
    • domains_gene_length.Rds: Rds file with domain lengths of A-domains of Basidiobolus NRPS proteins
    • domains_species.Rds: Rds files with coordinates of A-domains of Basidiobolus NRPS proteins
  • Fig7/ (Horizontal gene transfer in Basidiobolus species)
    • HGT_counts.txt: Summary of number of protein with matches to Bacterial protein sequences from RefSeq.
  • Fig8/ (Measurements from siderophore assays of Basidiobolus meristosporus CBS 931.37 and other treatments and control)
    • Siderophore_Measurements.txt: Data frame with raw easurements from the siderophore assays of Basidiobolus meristosporus CBS 931.37 , other treatments and control

Supplementary Figures

  • SF1/ (NRPS A-domain phylogenetic tree)
    • Bushley_names.txt: Data frame of names from the NRPS reconstruction of Bushley and Turgeon, 2010. Includes names of NRPS A-domains, Simplified names for NRPS A-domains and sub-family assignments.
    • NRPS.tre: Phylogenetic tree in NEWICK format from maximum likelihood reconstruction in RAxML using the predicted NRPS A-domains. Bootstrap supports from 1000 replicates are included as node.labels
  • SF4/ (PKS KS domain phylogenetic tree)
    • PKS.tre: Phylogenetic tree in NEWICK format from maximum likelihood reconstruction in RAxML using the predicted PKS KS domains. Bootstrap supports from 1000 replicates are included as node.labels
  • SF5/ (Summary of PKS domains in fungal species)
    • PKS_Fungi.Rds: Rds file with PKS domain counts for all fungal species included in the manuscript.
  • SF6/ (Summary of PKS domains in zygomycetes species)
    • PKS_counts.txt: Data frame with PKS domain counts for 69 zygomycete species included in the manuscript.
  • SF7/ (TC phylogenetic tree)
    • TC.tre: Phylogenetic tree in NEWICK format from maximum likelihood reconstruction in RAxML using the predicted TC core genes. Bootstrap supports from 1000 replicates are included as node.labels
  • SF8/ (HGT assay figures)
    • GFF_chrom.jcf7180000797043.Rds: Rds file with GFF from scaffold jcf7180000797043 from Basidiobolus heterosporus.
    • GFF_chrom.jcf7180000803233.Rds: Rds file with GFF from scaffold jcf7180000803233 from Basidiobolus heterosporus.
    • jcf7180000797043_HGTplot.rds: Rds file/Data frame with HGT assignments and coverage for gene models harbored in scaffold jcf7180000797043 of Basidiobolus heterosporus
    • jcf7180000803233_HGTplot.rds: Rds file/Data frame with HGT assignments and coverage for gene models harbored in scaffold jcf7180000803233 of Basidiobolus heterosporus
    • jcf7180000797043_Zscores.Rds: Rds file/Data frame with Z-scores for each position of scaffold jcf7180000797043 of Basidiobolus heterosporus
    • jcf7180000803233_Zscores.Rds: Rds file/Data frame with Z-scores for each position of scaffold jcf7180000803233 of Basidiobolus heterosporus
  • SF9/ (Intron number in HGT and fungal genes)
    • intron_number.Rds: Data frame with Pnumber of introns from genes in Basidiobolus meristosporus CBS 931.37 classified as product of HGT.
  • SF10/ (5-mer assay in HGT and fungal genes)
    • pentameres_simple.Rds: Rds file/data frame with the two main principal components for the 5-mer assay of Basidiobolus meristosporus CBS 931.37.
  • SF11/ (Codon usage assay in HGT and fungal genes)
    • pentameres_simple.Rds: Rds file/data frame with the two main principal components for the codon usage assay of Basidiobolus meristosporus CBS 931.37.

Additional files:

Required packages

library(ggplot2)
library(ggrepel)
library(ggpubr)
library(ggtree)
library(gridExtra)
library(phangorn)
library(phytools)
library(tidyverse)
library(reshape2)

Main Figures

Figures 3, 5 and 6 were constructed manually.

Figure 1. Summary of SM

zygo.all <- read.csv("Fig1/Zygo_SM_count.csv", header = T, stringsAsFactors = F)
zygo.all$SM_normalized <- zygo.all$SM/zygo.all$X..gene.models
zygo.all.m <- melt(zygo.all, id.vars = c("JGI.Identifier","Isolate.Species","Order","Phylum"))
zygo.all.m$gene_fam <- "Secondary_Metabolyte"
zygo.all.m$gene_fam[zygo.all.m$variable %in% c("Genome.size..Mb.")] <- "Genome_size"
zygo.all.m$gene_fam[zygo.all.m$variable %in% c("X..gene.models")] <- "Number_of_genes"
zygo.all.m$gene_fam[zygo.all.m$variable %in% c("SM")] <- 
"Secondary_Metabolyte_raw"
zygo.all.m$gene_fam[zygo.all.m$variable %in% c("SM_normalized")] <- "Proportion of SM per total genes"

# Plot
ggplot(zygo.all.m[!zygo.all.m$gene_fam %in% c("Secondary_Metabolyte_raw"),]) + geom_bar(aes(x=Isolate.Species, y=value, fill=variable), stat="identity") + 
  facet_grid(gene_fam ~ Order + Phylum, scales = "free", space = "free_x") + 
  scale_fill_viridis_d() + 
  theme_classic() + 
  theme(axis.text.x = element_text(angle = 90, hjust = 1))
## Warning: Removed 9 rows containing missing values (position_stack).

Figure 2. RNA expression

count.RNA <- readRDS("Fig2/count_RNA.Rds")

# Plot
ggplot(count.RNA, aes(x=counts.trans)) + geom_histogram(bins = 50) + 
      geom_point(data = count.RNA[!is.na(count.RNA$family),],aes(x=counts.trans, y=yval, colour=family, label = V1)) +    
      geom_text_repel(data = count.RNA[!is.na(count.RNA$family),],aes(x=counts.trans, y=yval, colour=family, label = V1)) +
  scale_x_continuous(breaks = c(0, 0.1, 1, 10, 100, 1000, 10000), trans = scales::log2_trans()) + theme_classic()  + xlab("Transcripts per million (TPM)") + ylab("Count") + scale_colour_viridis_d()
## Warning: Ignoring unknown aesthetics: label
## Warning: Transformation introduced infinite values in continuous x-axis

## Warning: Transformation introduced infinite values in continuous x-axis

## Warning: Transformation introduced infinite values in continuous x-axis
## Warning: Removed 2313 rows containing non-finite values (stat_bin).

Figure 4: A-domains

Reconstructing the domains

cols <- readRDS(file = "Fig4/Colors_arrow.Rds")
cols.origin <- readRDS(file = "Fig4/Colors_arrow_origin.Rds")
domains.species <- readRDS("Fig4/domains_species.Rds")
dom.gene.lenght <- readRDS("Fig4/domains_gene_length.Rds")

bashet.nrps <- ggplot(domains.species[[1]], aes(x=1)) + geom_segment(data = dom.gene.lenght[[1]], aes(x=0,xend=end,y=position, yend=position)) + geom_segment(aes(x=start,xend=finish,  y=position, yend=position, color=Class), arrow=arrow(length=unit(0.15,"cm"),  type = "closed"), size=2)  + theme_classic() + facet_grid(Gene ~ ., scales = "free")  + xlab("Position") + ylab("NRPS gene") + scale_color_manual(values = cols[names(cols) %in% unique(domains.species[[1]]$Class)]) + theme(axis.text.y=element_blank(),axis.ticks=element_blank(),legend.position = "bottom",strip.text.y = element_text(angle = 0), axis.line.y = element_blank()) + guides(fill=FALSE)

basmer.jgi.nrps <- ggplot(domains.species[[2]], aes(x=1)) + geom_segment(data = dom.gene.lenght[[2]], aes(x=0,xend=end,y=position, yend=position)) + geom_segment(aes(x=start,xend=finish,  y=position, yend=position, color=Class), arrow=arrow(length=unit(0.15,"cm"),  type = "closed"), size=2)  + theme_classic() + facet_grid(Gene ~ ., scales = "free")  + xlab("Position") + ylab("NRPS gene") + scale_color_manual(values = cols[names(cols) %in% unique(domains.species[[2]]$Class)]) + theme(axis.text.y=element_blank(),axis.ticks=element_blank(),legend.position = "bottom",strip.text.y = element_text(angle = 0), axis.line.y = element_blank()) + guides(fill=FALSE)

basmer.b9252.nrps <- ggplot(domains.species[[3]], aes(x=1)) + geom_segment(data = dom.gene.lenght[[3]], aes(x=0,xend=end,y=position, yend=position)) + geom_segment(aes(x=start,xend=finish,  y=position, yend=position, color=Class), arrow=arrow(length=unit(0.15,"cm"),  type = "closed"), size=2)  + theme_classic() + facet_grid(Gene ~ ., scales = "free")  + xlab("Position") + ylab("NRPS gene") + scale_color_manual(values = cols[names(cols) %in% unique(domains.species[[3]]$Class)]) + theme(axis.text.y=element_blank(),axis.ticks=element_blank(),legend.position = "bottom",strip.text.y = element_text(angle = 0), axis.line.y = element_blank()) + guides(fill=FALSE)

# Plot
grid.arrange(bashet.nrps, basmer.jgi.nrps, basmer.b9252.nrps)

Figure 7: HGT counts

HGT.counts <- read.table("Fig7/HGT_counts.txt", sep = "\t", header = T)

HGT.counts$Origin <- factor(HGT.counts$Origin, levels = c("a-proteobacteria","b-proteobacteria","d-proteobacteria","e-proteobacteria","g-proteobacteria","proteobacteria","firmicutes","actinobacteria","high GC Gram+","enterobacteria","planctomycetes","CFB group bacteria","GNS bacteria","verrucomicrobia","fusobacteria","cyanobacteria","chlamydias","mycoplasmas","aquificales","bacteria","euryarchaeotes","archaea"))
HGT.counts.prop <- HGT.counts
HGT.counts.prop$bashet <- HGT.counts$bashet/9331
HGT.counts.prop$basmer_B9252 <- HGT.counts$basmer_B9252/13273
HGT.counts.prop$basmer_HGT <- HGT.counts$basmer_HGT/16111

HGT.counts[order(HGT.counts$Origin, HGT.counts$Origin),]
HGT.counts.prop[order(HGT.counts.prop$Origin, HGT.counts$Origin),]

HGT.counts.m <- melt(HGT.counts.prop)

# Plot
ggplot(HGT.counts.m, aes(x=variable,y=value,fill=Origin)) + geom_bar(stat='identity', position = 'fill') + scale_fill_manual(values = c("#FFAAAA","#E37B7B","#D46A6A","#801515","#550000","#2B0000","#ED8229","#03D5D5","#198E8E","#674A33","#E9038F","#FFFF04","#ED5DBA","#72335B","#7F0855","#08AF13","#AA5704","#73239F","#1CA06E","#221617","#736058","#4E4E4E")) + theme_bw() + xlab("Taxon") + ylab("Normalized proportion of genes with HGT evidence ") + theme_classic()

Figure 8: Siderophore assay

side.meas <- read.table("Fig8/Siderophore_Measurements.txt", sep = "\t", stringsAsFactors = F, header = T)

# Plot
ggbarplot(data = side.meas, x = "Label", y="Area", fill="Label", add = "mean_se") + scale_fill_viridis_d(begin=0.9, end = 0.1) + theme(legend.position = "none")

Supplementary figures

Sup. Figs 2, 3 and 12 were constructed by hand

Sup. Figure 1: NRPS tree

zygo_tree <- read.tree("SF1/NRPS.tre")

# Reading assignments from Bushley 2010
table.names <- read.table("SF1/Bushley_names.txt", sep = "\t", header = T, stringsAsFactors = F)

# Assigning names
alin.names.org <- data.frame(names=zygo_tree$tip.label, group=table.names$Group[match(zygo_tree$tip.label,table.names$name.or.i.)], stringsAsFactors = F)

# Names_step1
names.step1 <- strsplit(alin.names.org$names, split = "_") %>% lapply(function (x) paste(x[1], x[2])) %>% unlist 
names.step1[grep(names.step1, pattern = "/")] <- grep(names.step1, pattern = "/", value = T) %>% strsplit(split = " ") %>% lapply(function (x) x[1]) %>% unlist
names.step1 <-  gsub(names.step1, pattern = " 1$", replacement = "_1") %>% gsub(pattern = " 2$", replacement = "_2") %>% gsub(perl = T, pattern = " \\d+$", replacement = "_JGI")
names_NRPS <- data.frame(tip.label= zygo_tree$tip.label, file= names.step1, stringsAsFactors = F)

# Separating by genus
names_NRPS$file <- strsplit(names_NRPS$file, split = " ", fixed = T) %>% lapply(function (x) x[1]) %>% unlist %>% gsub(pattern = "jgi\\|", replacement = "") %>% strsplit(split = "|", fixed = T) %>% lapply(function (x) x[1]) %>% unlist 

names_NRPS$genus <- names_NRPS$file

# JGI names
all.jgi <- read.csv('JGI_code_taxonomy.csv')
head(all.jgi)
##     Phylum_name     Subphylum_name        Class_name    Order_name
## 1    Ascomycota     Pezizomycotina   Dothideomycetes  Pleosporales
## 2 Basidiomycota    Agaricomycotina    Agaricomycetes   Polyporales
## 3 Basidiomycota    Agaricomycotina    Agaricomycetes   Polyporales
## 4  Mucoromycota     Mucoromycotina    Incertae_sedis     Mucorales
## 5  Mucoromycota     Mucoromycotina    Incertae_sedis     Mucorales
## 6 Basidiomycota Ustilaginomycotina Exobasidiomycetes Exobasidiales
##          Family_name    file                                Species
## 1       Dacampiaceae  Aaoar1      Aaosphaeria arxii CBS 175.79 v1.0
## 2        Meruliaceae  Abobi1      Abortiporus biennis CCBS 521 v1.0
## 3        Meruliaceae Abobie1 Abortiporus biennis CIRM-BRFM1778 v1.0
## 4 Cunninghamellaceae Chlpad1         Absidia padenii NRRL 2977 v1.0
## 5 Cunninghamellaceae Absrep1          Absidia repens NRRL 1336 v1.0
## 6  Cryptobasidiaceae  Acain1      Acaromyces ingoldii MCA 4198 v1.0
all.jgi$genus <- strsplit(as.character(all.jgi$Species), split = " ", fixed = T) %>% lapply(function (x) x[1])

# Merging the names
zygo.tree <- merge(names_NRPS, all.jgi,by = 'file', no.dups = T, sort = F)
zygo.tree <- zygo.tree[c(2,4)]
fungi.tree <- merge(names_NRPS, all.jgi,by = 'genus', no.dups = T, sort = F)
fungi.tree <- fungi.tree[c(2,4)]
fungi.tree <- fungi.tree[!duplicated(fungi.tree),]

fungi.all <- rbind(fungi.tree, zygo.tree)
fungi.all <- fungi.all[grep(names_NRPS$tip.label, pattern = "Basme2finSC",invert = T),]
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_NRPS$tip.label, pattern = "N161|Basme2finSC", value = T), "Phylum_name" = "BasMer"))
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_NRPS$tip.label, pattern = "N168", value = T), "Phylum_name" = "BasHet"))

# Plotting the tree
zygo_fort <- fortify(zygo_tree)
## Warning: `data_frame()` is deprecated as of tibble 1.1.0.
## Please use `tibble()` instead.
## This warning is displayed once every 8 hours.
## Call `lifecycle::last_warnings()` to see where this warning was generated.
## Warning: `mutate_()` is deprecated as of dplyr 0.7.0.
## Please use `mutate()` instead.
## See vignette('programming') for more help
## This warning is displayed once every 8 hours.
## Call `lifecycle::last_warnings()` to see where this warning was generated.
zygo_fort$bootstrap <- NA
zygo_fort$bootstrap[!zygo_fort$isTip] <- as.numeric(zygo_tree$node.label)
## Warning: NAs introduced by coercion
zygo_fort$bootstrap[zygo_fort$bootstrap < 70] <- NA

ggtree(zygo_fort, size=0.3) %<+% fungi.all + geom_tiplab(size=1, aes(color=Phylum_name)) + geom_text(aes(label=bootstrap), vjust=-.5, hjust=-.5, size=1) + theme(legend.position = "bottom") 
## Warning: Removed 1461 rows containing missing values (geom_text).

Figure was finalized by hand in Adobe Illustrator 2020 to color the missing tip labels, improve legibility and legend, and extend root.

Sup. Figure 4: PKS tree

zygo_tree <- read.tree("SF4/PKS.tre")

# Assigning names
alin.names.org <- data.frame(names=zygo_tree$tip.label, stringsAsFactors = F)

# Names_step1
names.step1 <- strsplit(alin.names.org$names, split = "_") %>% lapply(function (x) paste(x[1], x[2])) %>% unlist 
names.step1[grep(names.step1, pattern = "/")] <- grep(names.step1, pattern = "/", value = T) %>% strsplit(split = " ") %>% lapply(function (x) x[1]) %>% unlist
names.step1 <-  gsub(names.step1, pattern = " 1$", replacement = "_1") %>% gsub(pattern = " 2$", replacement = "_2") %>% gsub(perl = T, pattern = " \\d+$", replacement = "_JGI")
names_NRPS <- data.frame(tip.label= zygo_tree$tip.label, file= names.step1, stringsAsFactors = F)

# Separating by genus
names_NRPS$file <- strsplit(names_NRPS$file, split = " ", fixed = T) %>% lapply(function (x) x[1]) %>% unlist %>% gsub(pattern = "jgi\\|", replacement = "") %>% strsplit(split = "|", fixed = T) %>% lapply(function (x) x[1]) %>% unlist 

names_NRPS$genus <- names_NRPS$file

# JGI names
all.jgi <- read.csv('JGI_code_taxonomy.csv')
all.jgi$genus <- strsplit(as.character(all.jgi$Species), split = " ", fixed = T) %>% lapply(function (x) x[1])

# Merging the names
zygo.tree <- merge(names_NRPS, all.jgi,by = 'file', no.dups = T, sort = F)
zygo.tree <- zygo.tree[c(2,4)]
fungi.tree <- merge(names_NRPS, all.jgi,by = 'genus', no.dups = T, sort = F)
fungi.tree <- fungi.tree[c(2,4)]

fungi.all <- rbind(zygo.tree, fungi.tree)
fungi.all <- fungi.all[grep(names_NRPS$tip.label, pattern = "Basme2finSC",invert = T),]
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_NRPS$tip.label, pattern = "N161|Basme2finSC", value = T), "Phylum_name" = "BasMer"))
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_NRPS$tip.label, pattern = "N168", value = T), "Phylum_name" = "BasHet"))

# Plotting the tree
zygo_fort <- fortify(zygo_tree)
zygo_fort$bootstrap <- NA
zygo_fort$bootstrap[!zygo_fort$isTip] <- as.numeric(zygo_tree$node.label)
## Warning: NAs introduced by coercion
zygo_fort$bootstrap[zygo_fort$bootstrap < 70] <- NA

ggtree(zygo_fort, size=0.3) %<+% fungi.all + geom_tiplab(size=1, aes(color=Phylum_name)) + geom_text(aes(label=bootstrap), vjust=-.5, hjust=-.5, size=1) + theme(legend.position = "bottom") 
## Warning: Removed 650 rows containing missing values (geom_text).

Figure was finalized by hand in Adobe Illustrator 2020 to color the missing tip labels, improve legibility and legend,and extend root.

Sup. Figure 5: Fungal PKS

dom.df.m <- readRDS("SF5/PKS_Fungi.Rds")

# Plot
ggplot(dom.df.m, aes(x=variable, fill=value, y=AA)) +
  geom_tile(colour="black", show.legend = F) +
  theme_classic() +
  facet_grid(Phylum.name + Species ~ ., scale = "free", space = "free") +
  scale_x_discrete(position = "top") +
  xlab("Domain") + 
  theme(strip.text.y = element_text(size=9, face = "bold", angle = 360),strip.background = element_rect(size=0), axis.text.x = element_text(angle = 90, hjust = 1))

Sup. Figure 6: Zygo PKS

dom.df <- read.table("SF6/PKS_counts.txt", sep = "\t", header = T, comment.char = '')

# Plot
dom.df.m <- melt(dom.df)
## Using file, Species, Order, Phylum, AA, PKS_KR, source as id variables
ggplot(dom.df.m, aes(x=variable, fill=value, y=AA)) +
  geom_tile(colour="black", show.legend = F) +
  theme_classic() +
  facet_grid(source + Order + Species ~ ., scale = "free", space = "free") +
  scale_x_discrete(position = "top") +
  xlab("Domain") + 
  theme(strip.text.y = element_text(size=1, face = "bold", angle = 360),strip.background = element_rect(size=0), axis.text.x = element_text(angle = 90, hjust = 1)) +
  scale_fill_viridis_c(begin = 0.8, end = 0.8, na.value = "grey90")

Sup. Figure 7: TC tree

zygo_tree <- read.tree("SF7/TC.tree")

# Assigning names
alin.names.org <- data.frame(names=zygo_tree$tip.label, stringsAsFactors = F)

# Names_step1
names.step1 <- strsplit(alin.names.org$names, split = "_") %>% lapply(function (x) paste(x[1], x[2])) %>% unlist 
names.step1[grep(names.step1, pattern = "/")] <- grep(names.step1, pattern = "/", value = T) %>% strsplit(split = " ") %>% lapply(function (x) x[1]) %>% unlist
names.step1 <-  gsub(names.step1, pattern = " 1$", replacement = "_1") %>% gsub(pattern = " 2$", replacement = "_2") %>% gsub(perl = T, pattern = " \\d+$", replacement = "_JGI")
names_TC <- data.frame(tip.label= zygo_tree$tip.label, file= names.step1, stringsAsFactors = F)

# Separating by genus
names_TC$file <- strsplit(names_TC$file, split = " ", fixed = T) %>% lapply(function (x) x[1]) %>% unlist %>% gsub(pattern = "jgi\\|", replacement = "") %>% strsplit(split = "|", fixed = T) %>% lapply(function (x) x[1]) %>% unlist 

names_TC$genus <- names_TC$file

# JGI names
all.jgi <- read.csv('JGI_code_taxonomy.csv')
all.jgi$genus <- strsplit(as.character(all.jgi$Species), split = " ", fixed = T) %>% lapply(function (x) x[1])

# Merging the names
zygo.tree <- merge(names_TC, all.jgi,by = 'file', no.dups = T, sort = F)
zygo.tree <- zygo.tree[c(2,4)]
fungi.tree <- merge(names_TC, all.jgi,by = 'genus', no.dups = T, sort = F)
fungi.tree <- fungi.tree[c(2,4)]

fungi.all <- rbind(zygo.tree, fungi.tree)
fungi.all <- fungi.all[grep(names_TC$tip.label, pattern = "Basme2finSC",invert = T),]
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_TC$tip.label, pattern = "N161|Basme2finSC", value = T), "Phylum_name" = "BasMer"))
fungi.all <- rbind(fungi.all, data.frame("tip.label" = grep(names_TC$tip.label, pattern = "N168", value = T), "Phylum_name" = "BasHet"))

# Plotting the tree
zygo_fort <- fortify(zygo_tree)
zygo_fort$bootstrap <- NA
zygo_fort$bootstrap[!zygo_fort$isTip] <- as.numeric(zygo_tree$node.label)
## Warning: NAs introduced by coercion
zygo_fort$bootstrap[zygo_fort$bootstrap < 70] <- NA

ggtree(zygo_fort, size=0.3) %<+% fungi.all + geom_tiplab(size=1, aes(color=Phylum_name)) + geom_text(aes(label=bootstrap), vjust=-.5, hjust=-.5, size=1) + theme(legend.position = "bottom") 
## Warning: Removed 1609 rows containing missing values (geom_text).

Figure was finalized by hand in Adobe Illustrator 2020 to color the missing tip labels, improve legibility and legend,and extend root.

Sup. Figure 8: HGT assay

  • Negative:
jcf7180000803233 <- readRDS(file = "SF8/jcf7180000803233_HGTplot.Rds")
chom.sub.all <- readRDS(file = "SF8/jcf7180000803233_Zscores.Rds")
GFF.chrom <- readRDS(file = "SF8/GFF_chrom.jcf7180000803233.Rds")

# Coverage plot
cov.plot <-ggplot(data = jcf7180000803233, aes(x=Pos, y=Cov)) + 
  geom_line() + 
  geom_segment(data = GFF.chrom, aes(x=Start, xend=End, y=-1, yend=-1, color=Origin),arrow=arrow(length=unit(0.1,"inches"),  type = "closed"), size = 1, lineend = "butt", linejoin = "mitre") + 
  scale_y_continuous(breaks=seq(0,1500,100)) + 
  theme_classic() + 
  theme(legend.position = "bottom")

# Z-score plot
zscore.plot <- ggplot(data=chom.sub.all, aes(x=Pos, y=zscore)) + 
  geom_point(size=0.5) + 
  geom_hline(yintercept=c(-1, 1), color="forestgreen") + 
  geom_hline(yintercept=c(-2, 2), color="red") + 
  geom_segment(data = GFF.chrom, aes(x=Start, xend=End, y=-1, yend=-1, color=Origin), size = 3) + 
  theme_bw()

# Plot
ggarrange(cov.plot, zscore.plot ,nrow = 2, labels = c("Coverage", "Zscore"))

  • Positive
jcf7180000797043 <- readRDS(file = "SF8/jcf7180000797043_HGTplot.Rds")
chom.sub.all <- readRDS(file = "SF8/jcf7180000797043_Zscores.Rds")
GFF.chrom <- readRDS(file = "SF8/GFF_chrom.jcf7180000797043.Rds")

# Coverage plot
cov.plot <-ggplot(data = jcf7180000797043, aes(x=Pos, y=Cov)) + 
  geom_line() + 
  geom_segment(data = GFF.chrom, aes(x=Start, xend=End, y=-1, yend=-1, color=Origin),arrow=arrow(length=unit(0.1,"inches"),  type = "closed"), size = 1, lineend = "butt", linejoin = "mitre") + 
  scale_y_continuous(breaks=seq(0,1500,100)) + 
  theme_classic() + 
  theme(legend.position = "bottom")

# Z-score plot
zscore.plot <- ggplot(data=chom.sub.all, aes(x=Pos, y=zscore)) + 
  geom_point(size=0.5) + 
  geom_hline(yintercept=c(-1, 1), color="forestgreen") + 
  geom_hline(yintercept=c(-2, 2), color="red") + 
  geom_segment(data = GFF.chrom, aes(x=Start, xend=End, y=-1, yend=-1, color=Origin), size = 3) + 
  theme_bw()

# Plot
ggarrange(cov.plot, zscore.plot ,nrow = 2, labels = c("Coverage", "Zscore"))

Figure was finalized by hand in Adobe Illustrator 2020

Sup. Figures 9: Intron Number

intron.df <- readRDS("SF9/intron_number.Rds")

# Plot
ggviolin(intron.df, x="origin.x",y="num_introns", fill="origin.x", add = "boxplot", add.params = list(fill = "white"), draw_quantiles = T) + 
  scale_fill_viridis_d(begin = 0.5) + 
  xlab("Presumptive origin") +
  ylab("Number of introns")
## Warning in regularize.values(x, y, ties, missing(ties)): collapsing to unique
## 'x' values

Sup. Figure 10: Pentamers (Local)

pca.penta <- readRDS(file = "SF10/pentameres_simple.Rds")
pca.penta.bg <- pca.penta[,c(1:3)]

# Plot
penta.p <- ggplot(pca.penta, aes(x=PC1, y=PC2, color=label)) + 
  geom_point(data=pca.penta.bg, color="grey90", alpha = .9) + 
  geom_point() +
  theme_bw() +
  facet_grid(rows = vars(label), cols=vars(origin)) + 
  scale_color_viridis_d(option = "magma", begin = 0, end = 0.8) +
  guides(fill=FALSE, color=FALSE)

penta.p

Sup. Figure 11. Codon Usage

pca.df <- readRDS(file = "SF11/codon_usage_simple.Rds")
pca.df.bg <- pca.df[,c(1:3)]

# Plot
codon.p <- ggplot(pca.df, aes(x=PC1, y=PC2, color=label)) + 
  geom_point(data=pca.df.bg, color="grey90", alpha = .9) + 
  geom_point() +
  theme_bw() +
  facet_grid(rows = vars(label), cols=vars(origin)) + 
  scale_color_viridis_d(option = "magma", begin = 0, end = 0.8) +
  guides(fill=FALSE, color=FALSE)

codon.p

Revision folder:

The revision folder includes the process of review suggested by G3. The only change in the data analysis was to determine the number of HGT candidate genes that are in scaffolds with no fungal genes. The process and subsequent results are found in the HGT_Revision.Rmd file